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拮抗细菌FD6分离自福建闽侯青口青菜根围土壤,采用凹玻片法和离体叶片接种法测定菌株FD6的抑菌能力,经16S rDNA序列比对和相关生理生化性状分析,对生防细菌FD6进行了鉴定,并通过PCR扩增、薄层层析、薄层色谱生物自显影等探讨FD6产生抗生素的种类及其抑菌效果。结果表明,FD6的细菌悬浮液可显著抑制灰霉病菌分生孢子萌发,抑制率达99%,FD6培养滤液的抑制率仅为31%;细菌悬浮液对番茄灰霉病防效达59.7%。FD6的16S rDNA序列与假单胞菌属Pseudomonas的相似性达99%,系统进化树显示与荧光假单胞菌P.fluorescens遗传距离最近,结合生理生化表型特征将FD6菌株鉴定为荧光假单胞菌P.fluorescens。菌株FD6可产生硝吡咯菌素、2,4-二乙酰基间苯三酚、藤黄绿脓菌素、嗜铁素、氢氰酸和蛋白酶等抗菌物质,不产生吩嗪-1-羧酸,其中,硝吡咯菌素可直接抑制番茄灰霉病菌孢子萌发和菌丝的生长。
The antagonistic bacterium FD6 was isolated from the soil in the root-cabbage soil of Minhouchoukou, Minhou, Fujian Province. The antibacterial ability of strain FD6 was determined by concave-slide method and inoculated leaf inoculation method. The results of 16S rDNA sequence analysis and physiological- Bacteria FD6 were identified, and by PCR amplification, thin layer chromatography, thin layer chromatography autoradiography FD6 to explore the type of antibiotics and antibacterial effect. The results showed that the bacterial suspension of FD6 could significantly inhibit the spore germination of Botrytis cinerea, the inhibition rate was 99% and the inhibition rate of FD6 culture filtrate was only 31%. The bacterium suspension against tomato Botrytis was 59.7%. The 16S rDNA sequence of FD6 shared 99% similarity with Pseudomonas, and the phylogenetic tree showed that the genetic distance from P.fluorescens was the closest. The FD6 strain was identified as a fluorescent pseudo-single Bacillus P. fluorescens. Strain FD6 produces pyrrolnitrin, 2,4-diacetylphloroglucinol, pyocyanin, pyritin, hydrocyanic acid and protease and other antibacterial substances, does not produce phenazine-1-carboxylic acid Among them, pyrrolnitrin could directly inhibit spore germination and mycelial growth of Botrytis cinerea.